Structural Biology & Engineering Laboratory

Structural Biology & Engineering Laboratory

​​Research in the ​Structural Biology & Engineering Laboratory is dedicated to elucidating the molecular basis of the function and (de)regulation of proteins central to cellular signalling networks. The specificity, adaptability and robustness of signalling networks arises from multiple transient interactions among multidomain proteins, often embedded within 'signalosomes'. The intermediate size and dynamic nature of the proteins involved often precludes conventional three-dimensional (3D) structural analysis. Our investigations are therefore based on an integrated structural biology approach that combines functional and structural data from multiple sources (see 'Main Techniques' below). Using these approaches we investigate in particular kinase-mediated signalling networks. 

One major focus of the laboratory is on the structure and function of Focal adhesion (FA)–associated proteins. FAs are large macromolecular assemblies through which the actin cytoskeleton is connected to the extracellular matrix (ECM). More than just anchoring the cell, FAs encode the state of the ECM into intracellular biochemical pathways that control cell morphology, migration, differentiation, proliferation, and survival. We have recently determined the molecular basis for site-specific activation of full-length FA kinase (FAK) (Brami-Cherrier, 2014; see Figure). We now use this 3D molecular framework of FAK to understand how the close FAK homologue Pyk2 has evolved the unique capacity of calcium-sensing. Both FAK and Pyk2 have elicited great interest because of their important roles in human health (including cancer invasiveness, heart dysfunction and neurodegeneration). We will therefore also pursue opportunities for medical intervention against erroneous activation of these molecules through targeted intramolecular or intermolecular interaction inhibitors. 

Facilities

  • Liquid dispenser Brand: Formulatrix,
    Model: Mantis
    Liquid dispenser Mantis is a low dead-volume, non-contact liquid dispenser perfect for automated PCR assay setup. Mantis can quickly and accurately dispense liquid in a 96- or 384-well plate format. Capable of dispensing as little as 0.1 uL, Mantis allows to reduce reaction volumes.
  • Protein Stability Analysis Brand: Harbinger Biotechnology,
    Model: Stargazer-2
    Protein Stability Analysis The label-free DSLS (differential static light scattering) method measures the aggregation of protein in a 384-well microplate under controlled thermal conditions. This thermal aggregation assay can be used for many applications such as thermal stability analysis of proteins using temperature scanning & isothermal methods or comparison of ligand specificity, including substrates, co-factors, inhibitors, etc…
  • Plate Reader Brand: BMG Labtech,
    Model: PHERAstar FS
    Plate Reader The PHERAstar FS is the ultimate multi-mode reader for high-throughput screening, combining the highest sensitivity with the fastest read times. Thanks to the unique Optic Module system, Simultaneous Dual Emission, UV/Vis spectrometer, and TRF Lasers, it allows a wide-range of measurements.
  • Microscale Thermophoresis (MST) Brand: Nanotemper Technologies,
    Model: Monolith NT.115
    Microscale Thermophoresis (MST) MicroScale Thermophoresis is the directed movement of particles in a microscopic temperature gradient. Any change of the hydration shell of biomolecules due to changes in their structure/conformation results in a relative change of the movement along the temperature gradient and is used to determine binding affinities.